Analysis of differentially methylated genes in CMML samples with and without <i>TET2</i> mutations.

<p><b>A</b>) Hierarchical cluster analysis based on abnormally methylated genes between CMML samples with <i>TET2</i> mutations and CMML samples without <i>TET2</i> mutations. B values are depicted using a pseudocolor scale (Red = Genes hypermethylated; Green = Genes hypomethylated). Samples are color coded. The top bar beneath the dendrogram refers CMML or healthy donor samples, second bar indicates CMML samples with or without <i>TET2</i> mutations and lower bar indicates cytogenetic risk of CMML patients. <b>B</b>) Box plots for hypermethylated genes in CMML samples with <i>TET2</i> mutations (<i>TET2</i>-mut) with respect to CMML <i>TET2</i> wild type (<i>TET2</i>-wt) samples. <b>C</b>) Box plots for hypermethylated genes in CMML <i>TET2</i>-wt samples respect to CMML samples with <i>TET2</i>-mut.</p

Analysis of 5 hmC and 5 mC levels in genes hypermethylated in CMML TET2-mut in comparison to TET2-wt patients.

<p>The percentage of 5 hmC, 5 mC and ratio between 5 hmC/5 mC were measured in 8 CMML <i>TET2</i>-mut and 5 <i>TET2</i>-wt patient samples using qPCR. Two CpG located 5′upstream (−38 and −244 bp respectively) to the CpG analyzed in the methylation array in the case of <i>LAX1</i>; one upstream CpG (59 bp) and another downstream CpG (58 bp) to the CpG in the case of <i>SLC22A12</i> gene and one CpG downstream (5 bp) in the case of <i>VHL</i> gene were analyzed. Median values of percentage of 5 hmC, 5 mC or ratio between 5 hmC/5 mC are indicated and P values were obtained using the 2-tailed T test or U Mann Whitney test. CG: CpG dinucleotide included in the array; CCGG: CpG dinucleotide in which 5 hmC and 5 mC have been analysed and TSS: transcriptional start site.</p